Lettuce variety solan

ABSTRACT

The present invention provides novel lettuce cultivar Solan and plant parts, seed, and tissue culture therefrom. The invention also provides methods for producing a lettuce plant by crossing the lettuce plants of the invention with themselves or another lettuce plant. The invention also provides lettuce plants produced from such a crossing as well as plant parts, seed, and tissue culture therefrom.

FIELD OF THE INVENTION

This invention is in the field of lettuce plants.

BACKGROUND OF THE INVENTION

The present invention relates to a lettuce (Lactuca sativa L.) varietydesignated Solan.

Practically speaking, all cultivated forms of lettuce belong to thehighly polymorphic species Lactuca sativa that is grown for its ediblehead and leaves. Lactuca sativa is in the Cichoreae tribe of theAsteraceae (Compositae) family. Lettuce is related to chicory,sunflower, aster, dandelion, artichoke, and chrysanthemum. Sativa is oneof about 300 species in the genus Lactuca. There are seven differentmorphological types of lettuce. The crisphead group includes the icebergand batavian types. Iceberg lettuce has a large, firm head with a crisptexture and a white or creamy yellow interior. The batavian lettucepredates the iceberg type and has a smaller and less firm head. Thebutterhead group has a small, soft head with an almost oily texture. Theromaine, also known as cos lettuce, has elongated upright leaves forminga loose, loaf-shaped head and the outer leaves are usually dark green.Leaf lettuce comes in many varieties, none of which form a head, andinclude the green oak leaf variety. Latin lettuce looks like a crossbetween romaine and butterhead. Stem lettuce has long, narrow leaves andthick, edible stems. Oilseed lettuce is a type grown for its large seedsthat are pressed to obtain oil. Latin lettuce, stem lettuce, and oilseedlettuce are seldom seen in the United States.

Presently, there are over one thousand known lettuce cultivars. As acrop, lettuce is grown commercially wherever environmental conditionspermit the production of an economically viable yield.

Lettuce, in general, and leaf lettuce in particular, is an important andvaluable vegetable crop. Thus, there is an ongoing need for improvedlettuce varieties.

SUMMARY OF THE INVENTION

According to the invention, there is provided a novel lettuce cultivardesignated Solan, also known as LS15618, which is a blonde type butterhead lettuce and has characteristics including blonde leaf color, highresistances to Downy Mildew (Bremia lactucae 16-35:EU and 5-9US), TomatoBushy Stunt virus (TBSV), and Nasonovia ribisnigri biotype Nr: 0, andintermediate resistance to Lettuce Mosaic Virus (LMV) strain Ls-1. Theinvention also encompasses the seeds of lettuce cultivar Solan, theplants of lettuce cultivar Solan, plant parts of the lettuce cultivarSolan (including leaves, seed, gametes), methods of producing seed fromlettuce cultivar Solan, and method for producing a lettuce plant bycrossing the lettuce cultivar Solan with itself or another lettuceplant, methods for producing a lettuce plant containing in its geneticmaterial one or more transgenes, and the transgenic lettuce plantsproduced by that method. The invention also relates to methods forproducing other lettuce plants derived from lettuce cultivar Solan andto lettuce plants, parts thereof and seed derived by the use of thosemethods. The present invention further relates to hybrid lettuce seedsand plants (and parts thereof including leaves) produced by crossinglettuce cultivar Solan with another lettuce plant.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of lettuce cultivar Solan. In embodiments, thetissue culture is capable of regenerating plants having all oressentially all of the physiological and morphological characteristicsof the foregoing lettuce plant and/or of regenerating plants having thesame or substantially the same genotype as the foregoing lettuce plant.In exemplary embodiments, the regenerable cells in such tissue culturesare meristematic cells, cotyledons, hypocotyl, leaves, pollen, embryos,roots, root tips, anthers, pistils, ovules, shoots, stems, petiole,pith, flowers, capsules and/or seeds as well as callus and/orprotoplasts derived from any of the foregoing. Still further, thepresent invention provides lettuce plants regenerated from the tissuecultures of the invention.

As a further aspect, the invention provides a method of producinglettuce seed, the method comprising crossing a plant of lettuce cultivarSolan with itself or a second lettuce plant. Optionally, the methodfurther comprises collecting the seed.

Another aspect of the invention provides methods for producing hybridsand other lettuce plants derived from lettuce cultivar Solan. Lettuceplants derived by the use of those methods are also part of theinvention as well as plant parts, seed, gametes and tissue culture fromsuch hybrid or derived lettuce plants.

In representative embodiments, a lettuce plant derived from lettucecultivar Solan comprises cells comprising at least one set ofchromosomes derived from lettuce cultivar Solan. In embodiments, alettuce plant or population of lettuce plants derived from lettucecultivar Solan comprises, on average, at least 6.25%, 12.5%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% of its alleles (i.e., theoretical allelic content; TAC) fromlettuce cultivar Solan, e.g., at least about 6.25%, 12.5%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% of the genetic complement of lettuce cultivar Solan, andoptionally is the result of a breeding process comprising one or twobreeding crosses and one or more of selfing, sibbing, backcrossingand/or double haploid techniques in any combination and any order. Inembodiments, the breeding process does not include a breeding cross, andcomprises selfing, sibbing, backcrossing and or double haploidtechnology. In embodiments, the lettuce plant derived from lettucecultivar Solan is one, two, three, four, five or more breeding crossesremoved from lettuce cultivar Solan.

In embodiments, a hybrid or derived plant from lettuce cultivar Solancomprises a desired added trait(s). In representative embodiments, alettuce plant derived from lettuce cultivar Solan comprises all of themorphological and physiological characteristics of lettuce cultivarSolan (e.g., as described in Tables 1 to 15, for example, blonde leafcolor, high resistance to Downy Mildew (Bremia lactucae 16-35:EU and5-9US), high resistance to TBSV, intermediate resistance to LMV strainLs-1 and/or high resistance to Nasonovia ribisnigri biotype Nr: 0. Inembodiments, the lettuce plant derived from lettuce cultivar Solancomprises essentially all of the morphological and physiologicalcharacteristics of lettuce cultivar Solan (e.g., as described in Tables1 to 15, for example, blonde leaf color, high resistances to DownyMildew (Bremia lactucae 16-35:EU and 5-9US), TBSV and/or Nasonoviaribisnigri biotype Nr: 0) and/or intermediate resistance to LMV strainLs-1, with the addition of a desired added trait(s).

The invention also relates to methods for producing a lettuce plantcomprising in its genetic material one or more transgenes and to thetransgenic lettuce plant produced by those methods (and progeny lettuceplants comprising the transgene). Also provided are plant parts, seedand tissue culture from such transgenic lettuce plants, optionallywherein one or more cells in the plant part, seed, or tissue culturecomprises the transgene. The transgene can be introduced via planttransformation and/or breeding techniques.

In another aspect, the present invention provides for single geneconverted plants of lettuce cultivar Solan. Plant parts, seed, andtissue culture from such single gene converted plants are alsocontemplated by the present invention. The single transferred gene maybe a dominant or recessive allele. In representative embodiments, thesingle transferred gene confers such traits as male sterility, herbicideresistance, pest resistance (e.g., insect and/or nematode resistance),modified fatty acid metabolism, modified carbohydrate metabolism,disease resistance (e.g., for bacterial, fungal and/or viral disease),male fertility, enhanced nutritional quality, improved appearance (e.g.,color), improved salt tolerance, industrial usage, or any combinationthereof. The single gene may be a naturally occurring lettuce gene or atransgene introduced into lettuce through genetic engineeringtechniques.

The invention further provides methods for developing lettuce plants ina lettuce plant breeding program using plant breeding techniquesincluding, for example, recurrent selection, backcrossing, pedigreebreeding, double haploid techniques, restriction fragment lengthpolymorphism enhanced selection, genetic marker enhanced selectionand/or transformation. Seeds, lettuce plants, and parts thereof,produced by such breeding methods are also part of the invention.

The invention also provides methods of multiplication or propagation oflettuce plants of the invention, which can be accomplished using anymethod known in the art, for example, via vegetative propagation and/orseed.

The invention further provides a method of producing food or feedcomprising (a) obtaining a lettuce plant of the invention, optionallywherein the plant has been cultivated to maturity, and (b) collecting atleast one lettuce plant or part thereof (e.g., leaves) from the plant.

Additional aspects of the invention include harvested products andprocessed products from the lettuce plants of the invention. A harvestedproduct can be a whole plant or any plant part, as described herein.Thus, in some embodiments, a non-limiting example of a harvested productincludes a seed, a leaf and/or a stem.

In representative embodiments, a processed product includes, but is notlimited to: cut, sliced, ground, pureed, dried, canned, jarred, washed,packaged, frozen and/or heated leaves and/or seeds of the lettuce plantsof the invention, or any other part thereof. In embodiments, a processedproduct includes a sugar or other carbohydrate, fiber, protein and/oraromatic compound that is extracted, purified or isolated from a lettuceplant of the invention. In embodiments, the processed product includeswashed and packaged leaves (or parts thereof) of the invention.

The seed of the invention can optionally be provided as an essentiallyhomogenous population of seed of a single plant or cultivar. Essentiallyhomogenous populations of seed are generally free from substantialnumbers of other seed, e.g., at least about 90%, 95%, 96%, 97%, 98% or99% pure.

In representative embodiments, the invention provides a seed of lettucecultivar Solan.

As a further aspect, the invention provides a plant of lettuce cultivarSolan.

As an additional aspect, the invention provides a lettuce plant, or apart thereof, having all or essentially all of the physiological andmorphological characteristics of a plant of lettuce cultivar Solan.

As another aspect, the invention provides leaves and/or seed of thelettuce plants of the invention and a processed product from the leavesand/or seed of the inventive lettuce plants.

As still another aspect, the invention provides a method of producinglettuce seed, the method comprising crossing a lettuce plant of theinvention with itself or a second lettuce plant. The invention alsoprovides seed produced by this method and plants produced by growing theseed.

As yet a further aspect, the invention provides a method for producing aseed of a lettuce plant derived from lettuce cultivar Solan, the methodcomprising: (a) crossing a lettuce plant of lettuce cultivar Solan witha second lettuce plant; and (b) allowing seed of a lettuce plant derivedfrom lettuce cultivar Solan to form. In embodiments, the method furthercomprises: (c) growing a plant from the seed derived from lettucecultivar Solan of step (b); (d) selfing the plant grown from the lettuceseed derived from lettuce cultivar Solan or crossing it to a secondlettuce plant to form additional lettuce seed derived from lettucecultivar Solan, and (e) repeating steps (c) and (d) 0 or more times togenerate further derived lettuce seed. Optionally, the method comprises:(e) repeating steps (c) and (d) one or more times (e.g., one to three,one to five, one to six, one to seven, one to ten, three to five, threeto six, three to seven, three to eight or three to ten times) togenerate further derived lettuce plants. As another option, the methodcan comprise collecting the seed. The invention also provides seedproduced by these methods and plants produced by growing the seed.

As another aspect, the invention provides a method of producing lettuceleaves, the method comprising: (a) obtaining a plant of lettuce cultivarSolan, optionally wherein the plant has been cultivated to maturity; and(b) collecting leaves from the plant. The invention also provides theleaves produced by this method.

Still further, as another aspect, the invention provides a method ofvegetatively propagating a plant of lettuce cultivar Solan. In anon-limiting example, the method comprises: (a) collecting tissuecapable of being propagated from a plant of lettuce cultivar Solan; (b)cultivating the tissue to obtain proliferated shoots; and (c) rootingthe proliferated shoots to obtain rooted plantlets. Optionally, theinvention further comprises growing plants from the rooted plantlets.The invention also encompasses the plantlets and plants produced bythese methods.

As an additional aspect, the invention provides a method of introducinga desired added trait into lettuce cultivar Solan, the methodcomprising: (a) crossing a first plant of lettuce cultivar Solan with asecond lettuce plant that comprises a desired trait to produce F1progeny; (b) selecting an F1 progeny that comprises the desired trait;(c) crossing the selected F1 progeny with lettuce cultivar Solan toproduce backcross progeny; and (d) selecting backcross progenycomprising the desired trait to produce a plant derived from lettucecultivar Solan comprising a desired trait. In embodiments, the selectedprogeny comprises all or essentially all the morphological andphysiological characteristics of the first plant of lettuce cultivarSolan. Optionally, the method further comprises: (e) repeating steps (c)and (d) one or more times in succession (e.g., one to three, one tofive, one to six, one to seven, one to ten, three to five, three to six,three to seven, three to eight or three to ten times) to produce a plantderived from lettuce cultivar Solan comprising the desired trait.

In representative embodiments, the invention also provides a method ofproducing a plant of lettuce cultivar Solan comprising a desired addedtrait, the method comprising introducing a transgene conferring thedesired trait into a plant of lettuce cultivar Solan. The transgene canbe introduced by transformation methods (e.g., genetic engineering) orbreeding techniques. In embodiments, the plant comprising the transgenecomprises all or essentially all of the morphological and physiologicalcharacteristics of lettuce cultivar Solan.

The invention also provides lettuce plants produced by the methods ofthe invention, wherein the lettuce plant has the desired added trait aswell as seed from such lettuce plants.

According to the foregoing methods, the desired added trait can be anysuitable trait known in the art including, for example, male sterility,male fertility, herbicide resistance, insect or pest (e.g., insectand/or nematode) resistance, modified fatty acid metabolism, modifiedcarbohydrate metabolism, disease resistance (e.g., for bacterial, fungaland/or viral disease), enhanced nutritional quality, increasedsweetness, increased flavor, improved ripening control, improved salttolerance, industrial usage, or any combination thereof.

In representative embodiments, a transgene conferring herbicideresistance confers resistance to glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinate, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine,benzonitrile, or any combination thereof.

In representative embodiments, a transgene conferring pest resistance(e.g., insect and/or nematode resistance) encodes a Bacillusthuringiensis endotoxin.

In representative embodiments, transgenic plants, transformed plants,hybrid plants and lettuce plants derived from lettuce cultivar Solanhave at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of the morphologicaland physiological characteristics of lettuce cultivar Solan (e.g., asdescribed in Tables 1 to 15) in any combination, for example, blondeleaf color, high resistances to Downy Mildew (Bremia lactucae 16-35:EUand 5-9US), TBSV and/or Nasonovia ribisnigri biotype Nr: 0 and/orintermediate resistance to LMV strain Ls-1, or even all of themorphological and physiological characteristics of lettuce cultivarSolan, so that said plants are not significantly different for saidtraits than lettuce cultivar Solan, as determined at the 5% significancelevel when grown in the same environmental conditions; optionally, withthe presence of one or more desired additional traits (e.g., malesterility, disease resistance, pest or insect resistance, herbicideresistance, and the like).

In embodiments, the plants of the invention have at least 1, 2, 3, 4, 5,6, 7, 8, 9, 10 or more of the morphological and physiologicalcharacteristics of lettuce cultivar Solan (e.g., as described in Tables1 to 15). For example, the plants of the invention can have one, two,three, four, five, or more (in any combination) or even all of thefollowing characteristics: blonde leaf color, high resistances to DownyMildew (Bremia lactucae 16-35:EU and 5-9US), TBSV and/or Nasonoviaribisnigri biotype Nr: 0 and/or intermediate resistance to LMV strainLs-1.

The invention also encompasses plant parts, plant material, pollen,ovules, leaves, fruit and seed from the lettuce plants of the invention.Also provided is a tissue culture of regenerable cells from the lettuceplants of the invention, where optionally, the regenerable cells are:(a) embryos, meristem, leaves, pollen, cotyledons, hypocotyls, roots,root tips, anthers, flowers, pistils, ovules, seed, shoots, stems,stalks, petioles, pith and/or capsules; or (b) callus or protoplastsderived from the cells of (a). Further provided are lettuce plantsregenerated from a tissue culture of the invention.

In still yet another aspect, the invention provides a method ofdetermining a genetic characteristic of lettuce cultivar Solan or aprogeny thereof, e.g., a method of determining a genotype of lettucecultivar Solan or a progeny thereof using molecular genetic techniques.In embodiments, the method comprises detecting in the genome of a Solanplant, or a progeny plant thereof, at least a first polymorphism, e.g.,comprises nucleic acid amplification and/or nucleic acid sequencing. Toillustrate, in embodiments, the method comprises obtaining a sample ofnucleic acids from the plant and detecting at least a first polymorphismin the nucleic acid sample (e.g., using one or more molecular markers).Optionally, the method may comprise detecting a plurality ofpolymorphisms (e.g., two or more, three or more, four or more, five ormore, six or more, eight or more or ten or more polymorphisms, etc.) inthe genome of the plant. In representative embodiments, the methodfurther comprises storing the results of the step of detecting thepolymorphism(s) on a computer readable medium. The invention furtherprovides a computer readable medium produced by such a method.

In addition to the exemplary aspects and embodiments described above,the invention is described in more detail in the description of theinvention set forth below.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, in part, on the development of a novellettuce cultivar having desirable characteristics including blonde leafcolor, high resistances to Downy Mildew (Bremia lactucae 16-35:EU and5-9US), TBSV and Nasonovia ribisnigri biotype Nr:0, and intermediateresistance to LMV strain Ls-1.

It should be appreciated that the invention can be embodied in differentforms and should not be construed as limited to the embodiments setforth herein. Rather, these embodiments are provided so that thisdisclosure will be thorough and complete, and will fully convey thescope of the invention to those skilled in the art.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. The terminology used in thedescription of the invention herein is for the purpose of describingparticular embodiments only and is not intended to be limiting of theinvention.

Unless the context indicates otherwise, it is specifically intended thatthe various features and embodiments of the invention described hereincan be used in any combination.

Moreover, the present invention also contemplates that in someembodiments of the invention, any feature or combination of features setforth herein can be excluded or omitted. To illustrate, if thespecification states that a composition comprises components A, B and C,it is specifically intended that any of A, B or C, or a combinationthereof, can be omitted and disclaimed singularly or in any combination.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. The terminology used in thedescription of the invention herein is for the purpose of describingparticular embodiments only and is not intended to be limiting of theinvention.

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

As used in the description of the invention and the appended claims, thesingular forms “a,” “an” and “the” are intended to include the pluralforms as well, unless the context clearly indicates otherwise.

As used herein, “and/or” refers to and encompasses any and all possiblecombinations of one or more of the associated listed items, as well asthe lack of combinations when interpreted in the alternative (“or”).

The term “about,” as used herein when referring to a measurable valuesuch as a dosage or time period and the like, is meant to encompassvariations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of thespecified amount.

The term “comprise,” “comprises” and “comprising” as used herein,specify the presence of the stated features, integers, steps,operations, elements, and/or components, but do not preclude thepresence or addition of one or more other features, integers, steps,operations, elements, components, and/or groups thereof.

As used herein, the transitional phrase “consisting essentially of”means that the scope of a claim is to be interpreted to encompass thespecified materials or steps recited in the claim “and those that do notmaterially affect the basic and novel characteristic(s)” of the claimedinvention. See, In re Herz, 537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463(CCPA 1976) (emphasis in the original); see also MPEP § 2111.03. Thus,the term “consisting essentially of” when used in a claim or thedescription of this invention is not intended to be interpreted to beequivalent to “comprising.”

“Allele”. An allele is any of one or more alternative forms of a gene,all of which relate to a trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

“Backcrossing”. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F1 with one of the parental genotype of the F1 hybrid.

“Big Vein virus”. Big vein is a disease of lettuce caused by LettuceMirafiori Big Vein Virus which is transmitted by the fungus Opidiumvirulentus, with vein clearing and leaf shrinkage resulting in plants ofpoor quality and reduced marketable value.

“Bolting”. The premature development of a flowering stalk, andsubsequent seed, before a plant produces a food crop. Bolting istypically caused by late planting when temperatures are low enough tocause vernalization of the plants.

“Bremialactucae”. An Oomycete that causes downy mildew in lettuce incooler growing regions.

“Core length”. Length of the internal lettuce stem measured from thebase of the cut and trimmed head to the tip of the stem.

“Corky root”. A disease caused by the bacterium Sphingomonassuberifaciens, which causes the entire taproot to become brown, severelycracked, and non-functional.

“Cotyledon”. One of the first leaves of the embryo of a seed plant;typically one or more in monocotyledons, two in dicotyledons, and two ormore in gymnosperms.

“Double haploid line”. A stable inbred line achieved by doubling thechromosomes of a haploid line, e.g., from anther culture. For example,some pollen grains (haploid) cultivated under specific conditionsdevelop plantlets containing 1n chromosomes. The chromosomes in theseplantlets are then induced to “double” (e.g., using chemical means)resulting in cells containing 2n chromosomes. The progeny of theseplantlets are termed “double haploid” and are essentially notsegregating any more (e.g., are stable). The term “double haploid” isused interchangeably herein with “dihaploid.”

“Essentially all of the physiological and morphologicalcharacteristics”. In embodiments, a plant having “essentially all of thephysiological and morphological characteristics” (and similar phrases)means a plant having the physiological and morphological characteristicsof a parent plant (e.g., a recurrent parent), except for thecharacteristics derived from a converted gene(s) (i.e., via backcrossingto a recurrent parent), a modified gene(s) resulting from gene editingtechniques, or an introduced transgene (i.e., introduced via genetictransformation techniques). In embodiments, a plant having “essentiallyall of the physiological and morphological characteristics” (and similarphrases) means a plant having all of the characteristics of thereference plant with the exception of five or fewer traits, 4 or fewertraits, 3 or fewer traits, 2 or fewer traits, or one trait. Inembodiments, a plant having “essentially all of the physiological andmorphological characteristics” (and similar phrases) optionally has ablonde leaf color, high resistances to Downy Mildew (Bremia lactucae16-35: EU and 5-9US), TBSV, and/or Nasonovia ribisnigri biotype Nr: 0and/or intermediate resistance to LMV strain Ls-1 (in any combination).

“First water date”. The date the seed first receives adequate moistureto germinate. This can and often does equal the planting date.

“Gene”. As used herein, “gene” refers to a segment of nucleic acidcomprising an open reading frame. A gene can be introduced into a genomeof a species, whether from a different species or from the same species,using transformation or various breeding methods.

“Head diameter”. Diameter of the cut and trimmed head, slicedvertically, and measured at the widest point perpendicular to the stem.

“Head height”. Height of the cut and trimmed head, sliced vertically,and measured from the base of the cut stem to the cap leaf.

“Head weight”. Weight of saleable lettuce head, cut and trimmed tomarket specifications.

“Inbred line”: As used herein, the phrase “inbred line” refers to agenetically homozygous or nearly homozygous population. An inbred line,for example, can be derived through several cycles of sib crossingand/or selfing and/or via double haploid production. In someembodiments, inbred lines breed true for one or more traits of interest.An “inbred plant” or “inbred progeny” is an individual sampled from aninbred line.

“Lettuce Mosaic virus”. A disease that can cause a stunted, deformed, ormottled pattern in young lettuce and yellow, twisted, and deformedleaves in older lettuce.

“Maturity date”. Maturity refers to the stage when the plants are offull size and/or optimum weight and/or in marketable form to be ofcommercial or economic value.

“Nasonovia ribisnigri”. A lettuce aphid that colonizes the innermostleaves of the lettuce plant, contaminating areas that cannot be treatedeasily with insecticides.

“Plant.” As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which plants can beregenerated, plant calli, plant clumps, and plant cells that are intactin plants or parts of plants, such as leaves, pollen, embryos,cotyledons, hypocotyl, roots, root tips, anthers, pistils, flowers,ovules, seeds, fruit, stems, and the like.

“Plant material”. The terms “plant material” and “material obtainablefrom a plant” are used interchangeably herein and refer to any plantmaterial obtainable from a plant including without limitation, leaves,stems, roots, flowers or flower parts, fruits, pollen, ovules, zygotes,seeds, cuttings, cell or tissue cultures, or any other part or productof the plant.

“Plant part”. As used herein, a “plant part” includes any part, organ,tissue or cell of a plant including without limitation an embryo,meristem, leaf, pollen, cotyledon, hypocotyl, root, root tip, anther,flower, flower bud, pistil, ovule, seed, shoot, stem, stalk, petiole,pith, capsule, a scion, a rootstock and/or a fruit including callus andprotoplasts derived from any of the foregoing.

“Quantitative Trait Loci”. Quantitative Trait Loci (QTL) refers togenetic loci that control to some degree, numerically representabletraits that are usually continuously distributed.

“Ratio of head height/diameter”. Head height divided by the headdiameter is an indication of the head shape; <1 is flattened, 1=round,and >1 is pointed.

“Regeneration”. Regeneration refers to the development of a plant fromtissue culture.

“Resistance”. As used herein the terms “resistance” and “tolerance” (andgrammatical variations thereof) are used interchangeably to describeplants that show reduced or essentially no symptoms to a specific biotic(e.g., a pest, pathogen or disease) or abiotic (e.g., exogenous orenvironmental, including herbicides) factor or stressor. In someembodiments, “resistant” or “tolerant” plants show some symptoms but arestill able to produce marketable product with an acceptable yield, e.g.,the yield may still be reduced and/or the plants may be stunted ascompared with the yield or growth in the absence of the biotic and/orabiotic factor or stressor. Those skilled in the art will appreciatethat the degree of resistance or tolerance may be assessed with respectto a plurality or even an entire field of plants. A lettuce plant may beconsidered “resistant” or “tolerant” if resistance/tolerance is observedover a plurality of plants (e.g., an average), even if particularindividual plants may be susceptible to the biotic or abiotic factor orstressor.

“RHS”. RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd., RHS Garden;Wisley, Woking; Surrey GU236QB, UK.

“Single gene converted”. A single gene converted or conversion plantrefers to a plant that is developed by plant breeding techniques (e.g.,backcrossing) or via genetic engineering wherein essentially all of thedesired morphological and physiological characteristics of a line arerecovered in addition to the single gene transferred into the line viathe plant breeding technique or via genetic engineering.

“Substantially equivalent characteristic”. A characteristic that, whencompared, does not show a statistically significant difference (e.g.,p=0.05) from the mean.

“Tip burn”. Means a browning of the edges or tips of lettuce leaves thatis a physiological response to alack of calcium.

“Tomato Bushy Stunt”. Also called “lettuce necrotic stunt”. A diseasethat causes stunting of growth and leaf mottling.

“Transgene”. A nucleic acid of interest that can be introduced into thegenome of a plant by genetic engineering techniques (e.g.,transformation) or breeding. The transgene can be from the same or adifferent species. If from the same species, the transgene can be anadditional copy of a native coding sequence or can present the nativesequence in a form or context (e.g., different genomic location and/orin operable association with exogenous regulatory elements such as apromoter) than is found in the native state. The transgene can comprisean open reading frame encoding a polypeptide or can encode a functionalnon-translated RNA (e.g., RNAi).

Botanical Description of the Lettuce Cultivar Solan (LS15618).

Characteristics. Lettuce cultivar Solan is a blonde type butter headlettuce variety. Lettuce variety Solan resulted from a cross betweenbutter head lettuce lines and subsequent numerous generations ofindividual plant selections chosen for blonde color, good upsideappearance with a nice head and good balance, disease resistances toDowny Mildew (BI: 16-35EU and 5-9US), LMV strain Ls-1 and resistance toNasonovia ribisnigri (aphid) biotype Nr:0.

Lettuce Solan has shown uniformity and stability for all traits, withinthe limits of environmental influence. It has been self-pollinated asufficient number of generations with careful attention to uniformity ofplant type. The variety has been increased with continued observationfor uniformity. No variant traits have been observed or are expected inlettuce cultivar Solan.

TABLE 1 Variety Description Information. Plant Type: Butter Seed Seedcolor: Black Light dormancy: Light not required Heat dormancy: Nottested Cotyledon to Fourth Leaf Stage Shape of cotyledons: IntermediateUndulation: Slight Anthocyanin distribution: Absent Rolling: PresentCupping: Slight Reflexing: Apical Margin Mature Leaves Margin incisiondepth: Absent/ shallow Margin indentation: Shallow dentate Marginundulation of the apical margin: Absent/ slight Green color of outerleaves: Medium green Anthocyanin distribution: Absent Glossiness: DullLeaf blistering: Absent/slight Trichomes: Absent Leaf thickness: ThickPlant at Market Stage Head shape: Spherical Head size class: Large Headweight (g): 579 Head firmness: Loose Core Diameter at base of head (cm):2.6 Core height from base of head to apex (cm): 3.1 Maturity (days)Spring: 60 to 75 days Summer: 52 to 65 days Autumn: 52 to 75 daysWinter: 90 to 118 days Adaptation Primary U.S. Regions of Adaptation(tested and proven adapted) Southwest (California, Arizona desert): YesWest Coast: Yes Southeast: Not tested Northeast: Not tested Greenhouse:Not tested Soil type: Both of mineral and organic Disease and StressReactions Virus Tomato Bushy Stunt virus (TBSV): Highly resistant Bigvein: Not tested Lettuce Mosaic virus: Not tested Cucumber Mosaic virus:Not tested Broad Bean Wilt: Not tested Turnip Mosaic virus: Not testedBest Western Yellows: Not tested Lettuce Infectious Yellows: Not testedFungal/Bacterial Bremia lactucae (Downy Mildew): Highly resistant to Bl:16-35EU and 5-9U5 Lettuce Mosaic Virus strain Ls-1: Intermediateresistance Powdery Mildew: Not tested Sclerotinia Rot: Not testedBacterial Soft Rot (Pseudomonas spp. & others): Not tested Botrytis(Gray Mold): Not tested Insects Cabbage Loopers: Not tested Nasonoviabiotype Nr:0: Highly resistant Root Aphids: Not tested Green PeachAphid: Not tested Physiological/Stress Tip burn: Intermediate resistanceHeat: Intermediate tolerance Drought: Not tested Cold: Not tested Salt:Not tested Brown Rib: Not tested Post-Harvest Pink Rib: Not testedRussett Spotting: Not tested Rusty Brown Discoloration: Not testedInternal Rib Necrosis (Blackheart, Gray Rib, Gray Streak): Not testedBrown Stain: Not tested

TABLE 2 Length (mm) of 4th Leaf at 20 Days Solan Lucan 48 33 44 46 38 3937 39 42 41 39 44 39 50 40 58 48 55 45 48 40 40 43 40 45 39 55 40 47 4554 48 46 48 48 47 49 50 50 49 ANOVA Response Variable: Length of 4thLeaf at 20 Days Source of Variation df SS MS F P-value Variety 1 0.1 0.10.003 0.956 ANOVA shows no significant difference (p > 0.05) in thelength of the 4th true leaf measured in mm on 20 day old seedlings.

TABLE 3 Width (mm) of 4th Leaf at 20 Days Solan Lucan 25 20 20 20 20 1920 18 21 18 19 20 18 22 23 21 20 19 17 25 22 23 24 22 26 21 25 22 25 1925 26 24 18 23 22 20 23 22 20 Response Variable: ANOVA Width of 4th Leafat 20 Days Source of Variation df SS MS F P-value Variety 1 11.03 11.031.831 0.184 ANOVA shows no significant difference (p > 0.05) in thewidth of the 4th true leaf measured in mm on 20 days old seedlings.

TABLE 4 Length to Width Index of 4th Leaf at 20 Days Solan Lucan 1.921.65 2.20 2.30 1.90 2.05 1.85 2.17 2.00 2.28 2.05 2.20 2.17 2.27 1.742.76 2.40 2.89 2.65 1.92 1.82 1.74 1.79 1.82 1.73 1.86 2.20 1.82 1.882.37 2.16 1.85 1.92 2.67 2.09 2.14 2.45 2.17 2.27 2.45 ResponseVariable: Length: Width ANOVA Index of 4th Leaf at 20 Days Source ofVariation df SS MS F P-value Variety 1 0.12 0.1199 1.317 0.258 ANOVAshows no significant difference (p > 0.05) in length to width index ofthe 4th true leaf on 20 days old seedlings.

TABLE 5 Head Weight (g) at Harvest Maturity Solan Lucan Radian SolanLucan Radian Solan Lucan Radian Loc. 1 Loc. 1 Loc. 1 Loc. 2 Loc. 2 Loc.2 Loc. 3 Loc. 3 Loc. 3 601.3 500.5 520.0 833.0 359.0 684.0 518.0 525.0447.0 638.7 390.5 499.3 777.0 330.0 352.0 478.0 422.0 445.0 518.8 599.0481.3 698.0 441.0 479.0 621.0 323.0 312.0 474.1 620.6 507.5 491.0 523.0483.0 686.0 384.0 461.0 567.8 273.3 569.3 708.0 434.0 555.0 682.0 385.0326.0 279.0 470.0 455.2 662.0 387.0 680.0 478.0 456.0 470.0 613.6 395.5556.1 558.0 377.0 742.0 569.0 379.0 300.0 369.5 447.4 540.0 716.0 499.0613.0 450.0 364.0 323.0 551.1 306.9 435.2 685.0 451.0 668.0 522.0 378.0525.0 574.8 404.1 384.3 721.0 393.0 380.0 580.0 515.0 430.0 567.3 316.9424.2 534.1 700.0 473.2 471.0 457.2 562.4 414.4 326.6 371.2 748.8 331.8507.9 536.7 479.0 448.0 506.5 379.8 550.0 500.4 408.1 515.2 616.0 651.4364.5 650.0 367.1 480.0 Response ANOVA Variable: Head Weight (g) atHarvest Maturity Source of df SS MS F P-value Variation Variety 2 464165232083 23.411 <0.0001 *** Location 2 155173  77586  7.826  0.0006 ***Variety: 4 131021  32755  3.603 0.0084 ** Location Duncan Variety MeanGrouping Solan 579.173 a Radian 482.995 b Lucan 428.768 c ANOVA showssignificant differences (p <0.05) in head weight (g) at harvest maturityfor variety, location, and in the interaction between variety andlocation.

TABLE 6 Plant Height (cm) at Harvest Maturity Solan Lucan Radian SolanLucan Radian Solan Lucan Radian Loc. 1 Loc. 1 Loc. 1 Loc. 2 Loc. 2 Loc.2 Loc. 3 Loc. 3 Loc. 3 48.0 36.0 42.0 50.0 41.2 41.8 50.8 46.0 48.8 42.038.0 45.0 51.0 46.3 49.2 51.5 40.0 46.0 38.0 40.0 44.0 47.0 43.4 47.750.7 48.0 44.0 36.0 31.0 45.0 49.0 45.7 47.5 51.2 45.6 46.4 40.0 38.044.0 45.0 48.1 49.6 49.3 40.7 47.8 38.0 38.0 46.0 46.0 43.6 47.8 50.046.4 40.1 34.0 37.0 45.0 43.0 45.0 45.9 49.7 48.4 47.5 42.0 39.0 47.046.0 42.9 45.2 48.2 40.2 42.2 46.0 40.0 43.0 52.0 44.4 44.0 49.0 39.841.1 44.0 38.0 43.0 51.0 46.5 42.4 47.4 41.0 52.5 42.0 39.0 43.0 44.038.0 44.0 42.0 36.0 43.0 46.0 36.0 45.0 46.0 38.0 40.0 46.0 36.0 44.046.0 38.0 45.0 47.0 36.0 50.0 45.0 38.0 42.0 46.0 37.0 45.0 ResponseANOVA Variable: Plant Height (cm) at Harvest Maturity Source of df SS MSF P-value Variation Variety 2 608.9 304.4  32.61 <0.0001 *** Location 2693.7 346.8  37.15 <0.0001 *** Variety: 4 178.3 44.6   5.53  0.0004 ***Location Duncan Variety Mean Grouping Solan 45.895 a Radian 45.062 aLucan 40.755 b ANOVA shows significant differences (p <0.05) in plantheight (cm) at harvest maturity for variety, location, and in theinteraction between variety and location.

TABLE 7 Length of Frame Leaves (cm) at Harvest Maturity Solan LucanRadian Solan Lucan Radian Solan Lucan Radian Loc. 1 Loc. 1 Loc. 1 Loc. 2Loc. 2 Loc. 2 Loc. 3 Loc. 3 Loc. 3 16.0 14.0 15.0 17.6 17.3 15.5 17.816.2 16.0 14.0 14.0 12.0 17.3 23.1 14.8 16.9 18.3 12.8 18.0 14.0 14.017.4 15.2 17.8 16.9 16.5 15.9 16.0 15.0 13.0 17.9 19.0 17.1 19.4 15.013.0 13.0 17.0 14.0 16.4 18.2 16.9 18.0 14.8 15.7 14.0 13.0 16.0 17.519.4 25.2 17.6 16.5 15.1 13.0 15.0 12.0 18.0 18.7 16.0 17.8 16.0 15.015.0 18.0 14.0 17.8 17.8 15.5 17.4 15.8 15.4 16.0 16.0 12.0 17.0 18.015.6 18.4 16.0 13.3 18.0 15.0 16.0 18.5 17.7 16.7 17.8 15.8 15.8 14.017.0 12.0 18.0 12.0 12.0 15.0 15.0 16.0 16.0 15.0 16.0 17.0 16.0 18.019.0 16.0 13.0 14.0 13.0 15.0 17.0 16.0 16.0 14.0 17.0 18.0 19.0 14.013.0 Response ANOVA Variable: Length of Frame Leaves (cm) at HarvestMaturity Source of df SS MS F P-value Variation Variety 2 51.6  25.80 8.193 <0.0001 *** Location 2 138.3  69.15  21.957 <0.0001 *** Variety:4 23.9   5.98  1.964 0.1049   Location Duncan Variety Mean GroupingSolan 16.735 a Lucan 16.183 a Radian 15.153 b ANOVA shows significantdifferences (p <0.05) in length of frame leaves (cm) at harvest maturityfor variety and location, but there were no significant differences inthe interaction between variety and location.

TABLE 8 Width of Frame Leaves (cm) at Harvest Maturity Solan LucanRadian Solan Lucan Radian Solan Lucan Radian Loc. 1 Loc. 1 Loc. 1 Loc. 2Loc. 2 Loc. 2 Loc. 3 Loc. 3 Loc. 3 20.0 17.0 25.0 26.3 24.0 22.1 20.022.2 21.8 18.0 19.0 20.0 23.9 27.5 21.8 22.0 21.5 18.5 20.0 22.0 22.021.2 24.0 17.9 19.9 20.0 19.4 21.0 25.0 21.0 24.1 23.0 22.0 23.3 19.016.6 20.0 34.0 22.0 26.0 24.4 24.5 20.0 19.1 19.9 23.0 17.0 22.0 25.722.9 24.4 18.5 20.6 20.2 20.0 23.0 18.0 26.0 23.9 18.1 24.2 22.5 21.017.0 22.0 22.0 21.7 22.6 21.0 20.2 20.8 20.0 16.0 21.0 19.0 22.3 23.422.0 21.3 22.4 17.3 21.0 22.0 21.0 23.6 22.5 25.1 23.5 18.3 19.4 21.021.0 20.0 23.0 17.0 18.0 22.0 22.0 22.0 26.0 21.0 22.0 25.0 23.0 24.025.0 22.0 21.0 25.0 17.0 21.0 24.0 22.0 20.0 21.0 21.0 22.0 26.0 18.023.0 Response ANOVA Variable: Width of Frame Leaves (cm) at HarvestMaturity Source of df SS MS F P-value Variation Variety 2 31.9  15.94 2.646 0.0753    Location 2 124.8  62.38  10.355  <0.0001 *** Variety: 417.4   4.35  0.716 0.5830    Location ANOVA shows a significantdifference (p <0.05) in width of frame leaves (cm) at harvest maturityfor location, but there were no significant differences for variety andin the interaction between variety and location.

TABLE 9 Length to Width Index of Frame Leaves (cm) at Harvest MaturitySolan Lucan Radian Solan Lucan Radian Solan Lucan Radian Loc. 1 Loc. 1Loc. 1 Loc. 2 Loc. 2 Loc. 2 Loc. 3 Loc. 3 Loc. 3 0.80 0.82 0.60 0.670.72 0.70 0.89 0.73 0.73 0.78 0.74 0.60 0.72 0.84 0.68 0.77 0.85 0.690.90 0.64 0.64 0.82 0.63 0.99 0.85 0.83 0.82 0.76 0.60 0.62 0.74 0.830.78 0.83 0.79 0.78 0.65 0.50 0.64 0.63 0.75 0.69 0.90 0.77 0.79 0.610.76 0.73 0.68 0.85 1.03 0.95 0.80 0.75 0.65 0.65 0.67 0.69 0.78 0.880.74 0.71 0.71 0.88 0.82 0.64 0.82 0.79 0.74 0.86 0.76 0.77 1.00 0.760.63 0.76 0.77 0.71 0.86 0.71 0.77 0.86 0.68 0.76 0.78 0.79 0.67 0.760.86 0.81 0.67 0.81 0.60 0.78 0.71 0.67 0.68 0.68 0.73 0.62 0.71 0.730.68 0.70 0.75 0.76 0.73 0.62 0.56 0.76 0.71 0.71 0.73 0.80 0.67 0.810.82 0.73 0.78 0.57 Response Length to Width Index of Frame ANOVAVariable: Leaves (cm) at Harvest Maturity Source of df SS MS F P-valueVariation Variety 2 0.027 0.01349  1.85 0.1610    Location 2 0.1540.07694 10.57  0.0001 *** Variety: 4 0.064 0.01593  2.29 0.0646   Location ANOVA shows a significant difference (p <0.05) in length towidth index of frame leaves at harvest maturity stage for location, butthere were no significant differences for variety and the interactionbetween variety and location.

TABLE 10 Core Length (mm) from Base-Market Trimmed, Single Cap LeafSolan Lucan Radian Solan Lucan Radian Solan Lucan Radian Loc. 1 Loc. 1Loc. 1 Loc. 2 Loc. 2 Loc. 2 Loc. 3 Loc. 3 Loc. 3 25 10 20 30 27 35 48 3640 20 25 30 32 25 30 52 38 33 30 20 25 30 26 40 42 32 29 25 40 28 27 2938 43 27 45 25 20 35 30 30 41 40 31 39 20 20 20 28 31 40 40 38 29 20 2527 38 28 23 31 32 33 25 10 30 35 31 33 54 28 23 30 25 25 35 27 36 45 2646 25 35 30 32 22 22 46 33 39 15 10 20 20 25 25 35 25 30 30 35 10 35 3020 30 10 20 30 25 30 25 20 30 20 20 30 10 20 30 Response Core Length(mm) from Base- ANOVA Variable: Market Trimmed, Single Cap Leaf Sourceof df SS MS F P-value Variation Variety 2  588 294.2   6.85 0.0015 **Location 2 3474 1736.9   40.42 <0.0001 *** Variety: 4  483 120.7   3.010.0213 *  Location Duncan Variety Mean Grouping Solan 31.325 a Radian30.225 a Lucan 26.175 b ANOVA shows significant differences (p <0.05) incore length (mm) from base at harvest maturity for variety, location,and in the interaction between variety and location.

TABLE 11 Core Diameter (mm) from Base-Market Trimmed, Single Cap LeafSolan Lucan Radian Solan Lucan Radian Solan Lucan Radian Loc. 1 Loc. 1Loc. 1 Loc. 2 Loc. 2 Loc. 2 Loc. 3 Loc. 3 Loc. 3 20 10 25 29 25 30 28 2823 20 20 25 30 30 28 28 25 26 20 18 18 28 23 30 28 27 24 22 20 18 29 2930 29 20 25 23 22 18 29 26 31 30 28 28 20 20 20 30 28 33 25 28 20 23 2025 30 25 25 30 27 22 25 10 20 30 26 31 28 21 20 25 25 20 30 24 29 31 2720 25 28 22 31 24 27 30 30 25 20 10 15 15 25 20 25 20 20 24 30 10 25 2020 30 15 20 30 20 20 20 10 20 20 15 20 15 15 20 Response ANOVA Variable:Core Diameter (mm) Source of df SS MS F P-value Variation Variety 2256.5  128.3   9.32 <0.0001 *** Location 2 1514.6   757.3  55.02 <0.0001*** Variety: 4 114.6  28.7   2.166 0.0774   Location Duncan Variety MeanGrouping Solan 25.75 a Radian 23.08 b Lucan 22.35 b ANOVA showssignificant differences (p <0.05) in core length (mm) from base atharvest maturity for variety and location, but there was no significantdifferences in the interaction between variety and location.

TABLE 12 Core Length to Diameter Index from Base-Market Trimmed, SingleCap Leaf Solan Lucan Radian Solan Lucan Radian Solan Lucan Radian Loc. 1Loc. 1 Loc. 1 Loc. 2 Loc. 2 Loc. 2 Loc. 3 Loc. 3 Loc. 3 1.25 1.00 0.801.03 1.08 1.17 1.71 1.29 1.74 1.00 1.25 1.20 1.07 0.83 1.07 1.86 1.521.27 1.50 1.11 1.39 1.07 1.13 1.33 1.50 1.19 1.21 1.14 2.00 1.56 0.931.00 1.27 1.48 1.35 1.80 1.09 0.91 1.94 1.03 1.15 1.32 1.33 1.11 1.391.00 1.00 1.00 0.93 1.11 1.21 1.60 1.36 1.45 0.87 1.25 1.08 1.27 1.120.92 1.03 1.19 1.50 1.00 1.00 1.50 1.17 1.19 1.06 1.93 1.33 1.15 1.201.00 1.25 1.17 1.13 1.24 1.45 0.96 2.30 1.00 1.25 1.36 1.03 0.92 0.811.53 1.10 1.56 0.75 1.00 1.33 1.33 1.00 1.25 1.40 1.25 1.50 1.25 1.171.00 1.40 1.50 1.00 1.00 0.67 1.00 1.00 1.25 1.50 1.25 2.00 1.50 1.001.33 1.50 0.67 1.33 1.50 Response ANOVA Variable: Core Length toDiameter Index Source of df SS MS F P-value Variation Variety 2 0.4530.2264  3.564 0.0315 *  Location 2 1.911 0.9553  15.037 <0.0001 ***Variety: 4 0.593 0.1482  2.45 0.0503   Location Duncan Variety MeanGrouping Radian 1.32 a Solan 1.21 b Lucan 1.18 b ANOVA shows significantdifferences (p <0.05) in core length to diameter index at harvestmaturity for variety and location, but there was no significantdifferences in the interaction between variety and location.

TABLE 13 Height (cm) of Mature Seed Stalk Solan Lucan 31 28 28 28 27 2929 28 30 26 29 29 28 30 26 31 25 31 28 29 26 29 25 26 27 28 24 29 25 2824 31 23 27 25 27 25 28 27 28 25 30 ANOVA Response Variable: Source ofHeight (cm) of Mature Seed Stalk Variation df SS MS F P-value Variety 136.1 36.1 10.57 0.0024 ** Duncan Variety Mean Grouping Lucan 28.5 aSolan 26.6 b ANOVA shows a significant difference (p < 0.05) in theheight of mature seed stalk, and the average mature seed stalk height(cm) is 28.5 for Lucan and 26.6 for Solan.

TABLE 14 Spread (cm) of Mature Seed Stalk at Widest Point Solan Lucan 1113 11 22 8 15 9 12 9 12 10 13 8 15 11 14 9 12 7 12 10 13 8 14 12 13 1114 9 17 9 12 10 15 13 13 16 14 14 13 ANOVA Response Variable: Spread(cm) of Source of Mature Seed Stalk at Widest Point Variation df SS MS FP-value Variety 1 133.2 133.22 25.89 <0.0001 *** Duncan Variety MeanGrouping Lucan 13.9 a Solan 10.3 b ANOVA shows a significant difference(p < 0.05) in the spread of mature seed stalk, and the average matureseed stalk spread (cm) is 13.9 for Lucan and 10.3 for Solan.

TABLE 15 Height to Spread Index (cm) of Mature Seed Stalk Solan Lucan2.82 2.15 2.55 1.27 3.38 1.93 3.22 2.33 3.33 2.17 2.90 2.23 3.50 2.002.36 2.21 2.78 2.58 4.00 2.42 2.60 2.23 3.13 1.86 2.25 2.15 2.18 2.072.78 1.65 2.67 2.58 2.30 1.80 1.92 2.08 1.56 2.00 1.93 2.15 ANOVAResponse Variable: Source of Height to Spread Index (cm) Variation df SSMS F P-value Variety 1 3.782 3.782 16.41 0.0002 *** Duncan Variety MeanGrouping Solan 2.71 a Lucan 2.09 b ANOVA shows a significant difference(p < 0.05) in the height to spread index of mature seed stalk, and theaverage mature seed stalk height to spread index is 2.71 for Solan and2.09 for Lucan.

Further Embodiments of the Invention

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign nucleic acids including additional or modified versions ofnative (endogenous) nucleic acids (optionally driven by a non-nativepromoter) in order to alter the traits of a plant in a specific manner.Any nucleic acid sequences, whether from a different species or from thesame species, which are introduced into the genome using transformationor various breeding methods, are referred to herein collectively as“transgenes.” Over the last fifteen to twenty years, several methods forproducing transgenic plants have been developed, and in particularembodiments the present invention also relates to transformed versionsof lettuce plants disclosed herein.

Genetic engineering techniques can be used (alone or in combination withbreeding methods) to introduce one or more desired added traits intoplant, for example, lettuce cultivar Solan or progeny or lettuce plantsderived thereof.

Plant transformation generally involves the construction of anexpression vector that will function in plant cells. Optionally, such avector comprises one or more nucleic acids comprising a coding sequencefor a polypeptide or an untranslated functional RNA under control of, oroperatively linked to, a regulatory element (for example, a promoter).In representative embodiments, the vector(s) may be in the form of aplasmid, and can be used alone or in combination with other plasmids, toprovide transformed lettuce plants using transformation methods asdescribed herein to incorporate transgenes into the genetic material ofthe lettuce plant.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct nucleic acid transfermethod, such as microprojectile-mediated delivery (e.g., with abiolistic device), DNA injection, Agrobacterium-mediated transformation,electroporation, and the like. Transformed plants obtained from theplants (and parts and tissue culture thereof) of the invention areintended to be within the scope of this invention.

Expression Vectors for Plant Transformation—Selectable Markers.

Expression vectors typically include at least one nucleic acidcomprising or encoding a selectable marker, operably linked to aregulatory element (for example, a promoter) that allows transformedcells containing the marker to be either recovered by negativeselection, e.g., inhibiting growth of cells that do not contain theselectable marker, or by positive selection, e.g., screening for theproduct encoded by the selectable marker. Many commonly used selectablemarkers for plant transformation are well known in the transformationart, and include, for example, nucleic acids that code for enzymes thatmetabolically detoxify a selective chemical agent which may be anantibiotic or an herbicide, or nucleic acids that encode an alteredtarget which is insensitive to the inhibitor. Positive selection methodsare also known in the art.

One commonly used selectable marker for plant transformation is aneomycin phosphotransferase II (nptII) coding sequence, for example,isolated from transposon Tn5, which when placed under the control ofplant regulatory signals confers resistance to kanamycin. Fraley, etal., PNAS, 80:4803 (1983). Another commonly used selectable marker ishygromycin phosphotransferase, which confers resistance to theantibiotic hygromycin. Vanden Elzen, et al., Plant Mol. Biol., 5:299(1985).

Additional selectable markers of bacterial origin that confer resistanceto antibiotics include gentamycin acetyl transferase, streptomycinphosphotransferase, aminoglycoside-3′-adenyl transferase, the bleomycinresistance determinant. Hayford, et al., Plant Physiol., 86:1216 (1988);Jones, et al., Mol. Gen. Genet., 210:86 (1987); Svab, et al., Plant Mol.Biol., 14:197 (1990); Hille, et al., Plant Mol. Biol., 7:171 (1986).Other selectable markers confer resistance to herbicides such asglyphosate, glufosinate, or bromoxynil. Comai, et al., Nature,317:741-744 (1985); Gordon-Kamm, et al., Plant Cell, 2:603-618 (1990);and Stalker, et al., Science, 242:419-423 (1988).

Selectable markers for plant transformation that are not of bacterialorigin include, for example, mouse dihydrofolate reductase, plant5-enolpyruvylshikimate-3-phosphate synthase, and plant acetolactatesynthase. Eichholtz, et al., Somatic Cell Mol. Genet., 13:67 (1987);Shah, et al., Science, 233:478 (1986); and Charest, et al., Plant CellRep., 8:643 (1990).

Another class of selectable marker for plant transformation involvesscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance such as an antibiotic. These selectable markers areparticularly useful to quantify or visualize the spatial pattern ofexpression of a transgene in specific tissues and are frequentlyreferred to as a reporter gene because they can be fused to transgene orregulatory sequence for the investigation of nucleic acid expression.Commonly used reporters for screening presumptively transformed cellsinclude alpha-glucuronidase (GUS), alpha-galactosidase, luciferase andchloramphenicol, acetyltransferase. Jefferson, R. A., Plant Mol. Biol.,5:387 (1987); Teeri, et al., EMBO J., 8:343 (1989); Koncz, et al., PNAS,84:131 (1987); and DeBlock, et al., EMBO J., 3:1681 (1984).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissues are available. Molecular Probes,Publication 2908, IMAGENE GREEN, pp. 1-4 (1993) and Naleway, et al., J.Cell Biol., 115:151a (1991).

Green Fluorescent Protein (GFP) is also utilized as a marker for nucleicacid expression in prokaryotic and eukaryotic cells. Chalfie, et al.,Science, 263:802 (1994). GFP and mutants of GFP may be used asscreenable markers.

Expression Vectors for Plant Transformation—Promoters.

Transgenes included in expression vectors are generally driven by anucleotide sequence comprising a regulatory element (for example, apromoter). Numerous types of promoters are well known in thetransformation arts, as are other regulatory elements that can be usedalone or in combination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells.

Examples of promoters under developmental control include promoters thatpreferentially initiate transcription in certain tissues, such asleaves, roots, seeds, fibers, xylem vessels, tracheids, or sclerenchyma.Such promoters are referred to as “tissue-preferred.” Promoters thatinitiate transcription only in certain tissue are referred to as“tissue-specific.” A “cell type” specific promoter preferentially drivesexpression in certain cell types in one or more organs, for example,vascular cells in roots or leaves.

An “inducible” promoter is a promoter that is under environmentalcontrol. Examples of environmental conditions that may affecttranscription by inducible promoters include anaerobic conditions or thepresence of light. Tissue-specific, tissue-preferred, cell typespecific, and inducible promoters constitute the class of“non-constitutive” promoters. A “constitutive” promoter is a promoterthat is active under most environmental conditions.

A. Inducible Promoters:

An inducible promoter is operably linked to a nucleic acid forexpression in a plant. Optionally, the inducible promoter is operablylinked to a nucleotide sequence encoding a signal sequence which isoperably linked to a nucleic acid for expression in the plant. With aninducible promoter, the rate of transcription increases in response toan inducing agent.

Any inducible promoter can be used in the instant invention. See Ward,et al., Plant Mol. Biol., 22:361-366 (1993). Exemplary induciblepromoters include, but are not limited to, that from the ACEI systemwhich responds to copper (Melt, et al., PNAS, 90:4567-4571 (1993));promoter from the In2 gene from maize which responds tobenzenesulfonamide herbicide safeners (Hershey, et al., Mol. Gen.Genet., 227:229-237 (1991) and Gatz, et al., Mol. Gen. Genet., 243:32-38(1994)) or Tet repressor from Tn10 (Gatz, et al., Mol. Gen. Genet.,227:229-237 (1991)). A representative inducible promoter is a promoterthat responds to an inducing agent to which plants do not normallyrespond. An exemplary inducible promoter is the inducible promoter froma steroid hormone gene, the transcriptional activity of which is inducedby a glucocorticosteroid hormone. Schena, et al., PNAS, 88:0421 (1991).

B. Constitutive Promoters:

A constitutive promoter is operably linked to a nucleic acid forexpression in a plant or the constitutive promoter is operably linked toa nucleotide sequence encoding a signal sequence which is operablylinked to a nucleic acid for expression in a plant.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell, et al., Nature, 313:810-812 (1985)) and the promoters from suchgenes as rice actin (McElroy, et al., Plant Cell, 2:163-171 (1990));ubiquitin (Christensen, et al., Plant Mol. Biol., 12:619-632 (1989) andChristensen, et al., Plant Mol. Biol., 18:675-689 (1992)); pEMU (Last,et al., Theor. Appl. Genet., 81:581-588 (1991)); MAS (Velten, et al.,EMBO J., 3:2723-2730 (1984)) and maize H3 histone (Lepetit, et al., Mol.Gen. Genet., 231:276-285 (1992) and Atanassova, et al., Plant J., 2(3):291-300 (1992)). The ALS promoter, XbaI/NcoI fragment 5′ to theBrassica napus ALS3 structural gene (or a nucleotide sequence similarityto said XbaI/NcoI fragment), represents a particularly usefulconstitutive promoter. See PCT Application No. WO 96/30530.

C. Tissue-Specific or Tissue-Preferred Promoters:

A tissue-specific promoter is operably linked to a nucleic acid forexpression in a plant. Optionally, the tissue-specific promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a nucleic acid for expression in a plant.Plants transformed with a nucleic acid of interest operably linked to atissue-specific promoter transcribe the nucleic acid of interestexclusively, or preferentially, in a specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai, et al., Science, 23:476-482(1983) and Sengupta-Gopalan, et al., PNAS, 82:3320-3324 (1985)); aleaf-specific and light-induced promoter such as that from cab orrubisco (Simpson, et al., EMBO J., 4(11):2723-2729 (1985) and Timko, etal., Nature, 318:579-582 (1985)); an anther-specific promoter such asthat from LAT52 (Twell, et al., Mol. Gen. Genet., 217:240-245 (1989)); apollen-specific promoter such as that from Zm13 (Guerrero, et al., Mol.Gen. Genet., 244:161-168 (1993)) or a microspore-preferred promoter suchas that from apg (Twell, et al., Sex. Plant Reprod., 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments.

Transport of polypeptides produced by transgenes to a subcellularcompartment such as the chloroplast, vacuole, peroxisome, glyoxysome,cell wall, or mitochondrion, or for secretion into the apoplast, isgenerally accomplished by means of operably linking a nucleotidesequence encoding a signal sequence to the 5′ and/or 3′ region of anucleic acid encoding the polypeptide of interest. Signal sequences atthe 5′ and/or 3′ end of the coding sequence target the polypeptide toparticular subcellular compartments.

The presence of a signal sequence can direct a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker, et al., Plant Mol. Biol., 20:49 (1992); Close, P. S.,Master's Thesis, Iowa State University (1993); Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley,” Plant Mol. Biol., 9:3-17 (1987); Lerner, et al., PlantPhysiol., 91:124-129 (1989); Fontes, et al., Plant Cell, 3:483-496(1991); Matsuoka, et al., PNAS, 88:834 (1991); Gould, et al., J. Cell.Biol., 108:1657 (1989); Creissen, et al., Plant J, 2:129 (1991);Kalderon, et al., A short amino acid sequence able to specify nuclearlocation, Cell, 39:499-509 (1984); and Steifel, et al., Expression of amaize cell wall hydroxyproline-rich glycoprotein gene in early leaf androot vascular differentiation, Plant Cell, 2:785-793 (1990).

Foreign Polypeptide Transgenes and Agronomic Transgenes.

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign polypeptide then canbe extracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem., 114:92-6(1981).

According to a representative embodiment, the transgenic plant providedfor commercial production of foreign protein is a lettuce plant of theinvention. In another embodiment, the biomass of interest is seed. Forthe relatively small number of transgenic plants that show higher levelsof expression, a genetic map can be generated, for example viaconventional RFLP, PCR, and SSR analysis, which identifies theapproximate chromosomal location of the integrated DNA molecule. Forexemplary methodologies in this regard, see Methods in Plant MolecularBiology and Biotechnology, Glick and Thompson Eds., 269:284, CRC Press,Boca Raton (1993). Map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants, to determine if the latter have a commonparentage with the subject plant. Map comparisons can involvehybridizations, RFLP, PCR, SSR, and sequencing, all of which areconventional techniques.

Likewise, by means of the present invention, agronomic transgenes andother desired added traits can be expressed in transformed plants (andtheir progeny, e.g., produced by breeding methods). More particularly,plants can be genetically engineered to express various phenotypes ofagronomic interest or other desired added traits. Exemplary nucleicacids of interest in this regard conferring a desired added trait(s)include, but are not limited to, those categorized below:

A. Transgenes that Confer Resistance to Pests or Disease:

1. Plant disease resistance transgenes. Plant defenses are oftenactivated by specific interaction between the product of a diseaseresistance gene (R) in the plant and the product of a correspondingavirulence (Avr) gene in the pathogen. A plant line can be transformedwith a cloned resistance transgene to engineer plants that are resistantto specific pathogen strains. See, for example, Jones, et al., Science,266:789 (1994) (cloning of the tomato Cf-9 gene for resistance toCladosporium fulvum); Martin, et al., Science, 262:1432 (1993) (tomatoPto gene for resistance to Pseudomonas syringae pv. tomato encodes aprotein kinase); and Mindrinos, et al., Cell, 78:1089 (1994)(Arabidopsis RSP2 gene for resistance to Pseudomonas syringae).

2. A Bacillus thuringiensis protein, a derivative thereof, or asynthetic polypeptide modeled thereon. See, for example, Geiser, et al.,Gene, 48:109 (1986), who disclose the cloning and nucleotide sequence ofa Bt delta-endotoxin gene. Moreover, DNA molecules encodingdelta-endotoxin transgenes can be purchased from American Type CultureCollection, Manassas, Va., for example, under ATCC Accession Nos. 40098,67136, 31995, and 31998.

3. A lectin. See, for example, the disclosure by Van Damme, et al.,Plant Mol. Biol., 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin transgenes.

4. A vitamin-binding protein such as avidin. See, e.g., PCT ApplicationNo. US 93/06487. The application teaches the use of avidin and avidinhomologues as larvicides against insect pests.

5. An enzyme inhibitor, for example, a protease or proteinase inhibitor,or an amylase inhibitor. See, for example, Abe, et al., J. Biol. Chem.,262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor); Huub, et al., Plant Mol. Biol., 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I); and Sumitani,et al., Biosci. Biotech. Biochem., 57:1243 (1993) (nucleotide sequenceof Streptomyces nitrosporeus alpha-amylase inhibitor).

6. An insect-specific hormone or pheromone, such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock, et al., Nature, 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

7. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem., 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor) and Pratt, etal., Biochem. Biophys. Res. Comm., 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also, U.S. Pat. No. 5,266,317 toTomalski, et al., who disclose transgenes encoding insect-specific,paralytic neurotoxins.

8. An insect-specific venom produced in nature, by a snake, a wasp, etc.For example, see Pang, et al, Gene, 116:165 (1992), for disclosure ofheterologous expression in plants of a transgene coding for a scorpioninsectotoxic peptide.

9. An enzyme responsible for a hyper-accumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivative,or another non-protein molecule with insecticidal activity.

10. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase, and a glucanase, whether natural or synthetic. See PCTApplication No. WO 93/02197 in the name of Scott, et al., whichdiscloses the nucleotide sequence of a callase transgene. DNA moleculeswhich contain chitinase-encoding sequences can be obtained, for example,from the ATCC under Accession Nos. 39637 and 67152. See also, Kramer, etal., Insect Biochem. Mol. Biol., 23:691 (1993), who teach the nucleotidesequence of a cDNA encoding tobacco hornworm chitinase, and Kawalleck,et al., Plant Mol. Biol., 21:673 (1993), who provide the nucleotidesequence of the parsley ubi4-2 polyubiquitin transgene.

11. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella, et al., Plant Mol. Biol., 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess,et al., Plant Physiol., 104:1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

12. A hydrophobic moment peptide. See PCT Application No. WO 95/16776(disclosure of peptide derivatives of tachyplesin which inhibit fungalplant pathogens) and PCT Application No. WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance).

13. A membrane permease, a channel former, or a channel blocker. Forexample, see the disclosure of Jaynes, et al., Plant Sci., 89:43 (1993),of heterologous expression of a cecropin-beta, lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

14. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein transgene is derived,as well as by related viruses. See Beachy, et al., Ann. Rev.Phytopathol., 28:451 (1990). Coat protein-mediated resistance has beenconferred upon transformed plants against alfalfa mosaic virus, cucumbermosaic virus, tobacco streak virus, potato virus X, potato virus Y,tobacco etch virus, tobacco rattle virus, and tobacco mosaic virus. Id.

15. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. SeeTaylor, et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions, Edinburgh, Scotland (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

16. A virus-specific antibody. See, for example, Tavladoraki, et al.,Nature, 366:469 (1993), who show that transgenic plants expressingrecombinant antibody transgenes are protected from virus attack.

17. A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo-alpha-1,4-D-polygalacturonasesfacilitate fungal colonization and plant nutrient released bysolubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See Lamb,et al., Bio/technology, 10:1436 (1992). The cloning and characterizationof a transgene which encodes a bean endopolygalacturonase-inhibitingprotein is described by Toubart, et al., Plant J., 2:367 (1992).

18. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann, et al., Bio/technology, 10:305 (1992), have shownthat transgenic plants expressing the barley ribosome-inactivatingtransgene have an increased resistance to fungal disease.

19. A lettuce mosaic potyvirus (LMV) coat protein transgene introducedinto Lactuca sativa in order to increase its resistance to LMVinfection. See Dinant, et al., Mol. Breeding, 3:1, 75-86 (1997).

Any disease or present resistance transgenes, including thoseexemplified above, can be introduced into a lettuce plant of theinvention through a variety of means including but not limited totransformation and breeding.

B. Transgenes that Confer Resistance to an Herbicide:

Exemplary polynucleotides encoding polypeptides that confer traitsdesirable for herbicide resistance include acetolactate synthase (ALS)mutants that lead to herbicide resistance such as the S4 and/or Hramutations ((resistance to herbicides including sulfonylureas,imidazolinones, triazolopyrimidines, pyrimidinyl thiobenzoates);glyphosate resistance (e.g.,5-enol-pyrovyl-shikimate-3-phosphate-synthase (EPSPS) transgene,including but not limited to those described in U.S. Pat. Nos.4,940,935, 5,188,642, 5,633,435, 6,566,587, 7,674,598 as well as allrelated application; or the glyphosate N-acetyltransferase (GAT)transgene, described in Castle et al., Science, 2004, 304:1151-1154; andin U.S. Patent Application Publication Nos. 20070004912, 20050246798,and 20050060767)); glufosinate resistance (e.g., BAR; see e.g., U.S.Pat. No. 5,561,236); 2,4-D resistance (e.g., aryloxy alkanoatedioxygenase or AAD-1, AAD-12, or AAD-13), HPPD resistance (e.g.,Pseudomonas HPPD) and PPO resistance (e.g., fomesafen,acifluorfen-sodium, oxyfluorfen, lactofen, fluthiacet-methyl,saflufenacil, flumioxazin, flumiclorac-pentyl, carfentrazone-ethyl,sulfentrazone); a cytochrome P450 or variant thereof that confersherbicide resistance or tolerance to, inter alia, HPPD-inhibitingherbicides, PPO-inhibiting herbicides and ALS-inhibiting herbicides(U.S. Patent Application Publication No. 20090011936; U.S. Pat. Nos.6,380,465; 6,121,512; 5,349,127; 6,649,814; and 6,300,544; and PCTInternational Publication No. WO 2007/000077); dicamba resistance (e.g.,dicamba monoxygenase), and traits desirable for processing or processproducts such as high oil (e.g., U.S. Pat. No. 6,232,529); modified oils(e.g., fatty acid desaturase transgenes (U.S. Pat. No. 5,952,544; PCTInternational Publication No. WO 94/11516)); modified starches (e.g.,ADPG pyrophosphorylases (AGPase), starch synthases (SS), starchbranching enzymes (SBE), and starch debranching enzymes (SDBE)); andpolymers or bioplastics (e.g., U.S. Pat. No. 5,602,321;beta-ketothiolase, polyhydroxybutyrate synthase, and acetoacetyl-CoAreductase (Schubert et al., J. Bacteriol., 1988, 170:5837-5847)facilitate expression of polyhydroxyalkanoates (PHAs)).

In embodiments, the polynucleotide encodes a polypeptide conferringresistance to an herbicide selected from glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinate, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine, andbenzonitrile.

Any transgene conferring herbicide resistance, including thoseexemplified above, can be introduced into the lettuce plants of theinvention through a variety of means including, but not limited to,transformation (e.g., genetic engineering techniques) and crossing.

C. Transgenes that Confer or Contribute to a Value-Added Trait:

1. Increased iron content of the lettuce, for example, by introducinginto a plant a soybean ferritin transgene as described in Goto, et al.,Acta Horticulturae., 521, 101-109 (2000).

2. Decreased nitrate content of leaves, for example, by introducing intoa lettuce a transgene coding for a nitrate reductase. See, for example,Curtis, et al., Plant Cell Rep., 18:11, 889-896 (1999).

3. Increased sweetness of the lettuce by introducing a transgene codingfor monellin that elicits a flavor 100,000 times sweeter than sugar on amolar basis. See Penarrubia, et al., Bio/technology, 10:561-564 (1992).

4. Modified fatty acid metabolism, for example, by introducing into aplant an antisense sequence directed against stearyl-ACP desaturase toincrease stearic acid content of the plant. See Knultzon, et al., PNAS,89:2625 (1992).

5. Modified carbohydrate composition effected, for example, byintroducing into plants a transgene coding for an enzyme that alters thebranching pattern of starch. See Shiroza, et al., J. Bacteria, 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase transgene); Steinmetz, et al., Mol. Gen. Genet.,20:220 (1985) (nucleotide sequence of Bacillus subtilis levansucrasetransgene); Pen, et al., Bio/technology, 10:292 (1992) (production oftransgenic plants that express Bacillus lichenifonnis alpha-amylase);Elliot, et al., Plant Mol. Biol., 21:515 (1993) (nucleotide sequences oftomato invertase transgenes); Sogaard, et al., J. Biol. Chem., 268:22480(1993) (site-directed mutagenesis of barley alpha-amylase transgene);and Fisher, et al., Plant Physiol., 102:1045 (1993) (maize endospermstarch branching enzyme II).

Any transgene that confers or contributes a value-added trait, includingthose exemplified above, can be introduced into the lettuce plants ofthe invention through a variety of means including, but not limited to,transformation (e.g., genetic engineering techniques) and crossing.

D. Transgenes that Control Male-Sterility:

1. Introduction of a deacetylase transgene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT. See, e.g., International Publication WO 01/29237.

2. Introduction of various stamen-specific promoters. See, e.g.,International Publications WO 92/13956 and WO 92/13957.

3. Introduction of the barnase and the barstar transgenes. See, e.g.,Paul, et al., Plant Mol. Biol., 19:611-622 (1992).

Any transgene that controls male sterility, including those exemplifiedabove, can be introduced into the lettuce plants of the inventionthrough a variety of means including, but not limited to, transformation(e.g., genetic engineering techniques) and crossing.

Methods for Plant Transformation.

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation” inMethods in Plant Molecular Biology and Biotechnology, Glick and ThompsonEds., CRC Press, Inc., Boca Raton, pp. 89-119(1993).

A. Agrobacterium-Mediated Transformation.

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch, et al., Science, 227:1229 (1985); Curtis, et al., Journal ofExperimental Botany, 45:279, 1441-1449 (1994); Torres, et al., PlantCell Tissue and Organ Culture, 34:3, 279-285 (1993); and Dinant, et al.,Molecular Breeding, 3:1, 75-86 (1997). A. tumefaciens and A. rhizogenesare plant pathogenic soil bacteria which genetically transform plantcells. The Ti and Ri plasmids of A. tumefaciens and A. rhizogenes,respectively, carry genes responsible for genetic transformation of theplant. See, for example, Kado, C. I., Crit. Rev. Plant Sci., 10:1(1991). Descriptions of Agrobacterium vector systems and methods forAgrobacterium-mediated transgene transfer are provided by Gruber, etal., supra, Miki, et al., supra, and Moloney, et al., Plant Cell Rep.,8:238 (1989). See also, U.S. Pat. No. 5,591,616 issued Jan. 7, 1997.

B. Direct Transgene Transfer.

Several methods of plant transformation collectively referred to asdirect transgene transfer have been developed as an alternative toAgrobacterium-mediated transformation. A generally applicable method ofplant transformation is microprojectile-mediated transformation whereinDNA is carried on the surface of microprojectiles measuring 1 micron to4 micron. The expression vector is introduced into plant tissues with abiolistic device that accelerates the microprojectiles to speeds of 300m/s to 600 m/s which is sufficient to penetrate plant cell walls andmembranes. Russell, D. R., et al., Plant Cell Rep., 12 (3, January),165-169 (1993); Aragao, F. J. L., et al., Plant Mol. Biol., 20 (2,October), 357-359 (1992); Aragao, F. J. L., et al., Plant Cell Rep., 12(9, July), 483-490 (1993); Aragao, Theor. Appl. Genet., 93:142-150(1996); Kim, J., Minamikawa, T., Plant Sci., 117:131-138 (1996);Sanford, et al., Part. Sci. Technol., 5:27 (1987); Sanford, J. C.,Trends Biotech., 6:299 (1988); Klein, et al., Bio/technology, 6:559-563(1988); Sanford, J. C., Physiol. Plant, 7:206 (1990); Klein, et al.,Bio/technology, 10:268 (1992).

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang, et al., Bio/technology, 9:996 (1991).Alternatively, liposome and spheroplast fusion have been used tointroduce expression vectors into plants. Deshayes, et al., EMBO J.,4:2731 (1985) and Christou, et al., PNAS, 84:3962 (1987). Direct uptakeof DNA into protoplasts using CaCl.sub.2 precipitation, polyvinylalcohol, or poly-L-ornithine has also been reported. Hain, et al., Mol.Gen. Genet., 199:161 (1985) and Draper, et al., Plant Cell Physiol.,23:451 (1982). Electroporation of protoplasts and whole cells andtissues have also been described. Saker, M., Kuhne, T., BiologiaPlantarum, 40(4):507-514 (1997/98); Donn, et al., In Abstracts of VIIthInternational Congress on Plant Cell and Tissue Culture IAPTC, A2-38, p.53 (1990); D'Halluin, et al., Plant Cell, 4:1495-1505 (1992); andSpencer, et al., Plant Mol. Biol., 24:51-61 (1994). See also Chupean, etal., Bio/technology, 7:5, 503-508 (1989).

Following transformation of plant target tissues, expression of theabove-described selectable marker transgenes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic lettuce line. The transgenic lettuce line couldthen be crossed with another (non-transformed or transformed) line inorder to produce a new transgenic lettuce line. Alternatively, a genetictrait that has been engineered into a particular plant cultivar usingthe foregoing transformation techniques could be introduced into anotherline using traditional breeding (e.g., backcrossing) techniques that arewell known in the plant breeding arts. For example, a backcrossingapproach could be used to move an engineered trait from a public,non-elite inbred line into an elite inbred line, or from an inbred linecontaining a foreign transgene in its genome into an inbred line orlines which do not contain that transgene. As used herein, “crossing”can refer to a simple X by Y cross, or the process of backcrossing,depending on the context.

Gene Conversions.

When the term “lettuce plant” is used in the context of the presentinvention, this term also includes any gene conversions of that plant orvariety. The term “gene converted plant” as used herein refers to thoselettuce plants that are developed, for example, by backcrossing, geneticengineering and/or mutation, wherein essentially all of the desiredmorphological and physiological characteristics of a variety arerecovered in addition to the one or more genes transferred into thevariety. To illustrate, backcrossing methods can be used with thepresent invention to improve or introduce a characteristic into thevariety. The term “backcrossing” as used herein refers to the repeatedcrossing of a hybrid progeny back to the recurrent parent, e.g.,backcrossing 1, 2, 3, 4, 5, 6, 7, 8, 9, or more times to the recurrentparent. The parental plant that contributes the gene for the desiredcharacteristic is termed the “nonrecurrent” or “donor parent.” Thisterminology refers to the fact that the nonrecurrent parent is generallyused one time in the breeding e.g., backcross) protocol and thereforedoes not recur. The gene that is transferred can be a native gene, amutated native gene or a transgene introduced by genetic engineeringtechniques into the plant (or ancestor thereof). The parental plant intowhich the gene(s) from the nonrecurrent parent are transferred is knownas the “recurrent” parent as it is used for multiple rounds in thebackcrossing protocol. Poehlman & Sleper (1994) and Fehr (1993). In atypical backcross protocol, the original variety of interest (recurrentparent) is crossed to a second variety (nonrecurrent parent) thatcarries the gene(s) of interest to be transferred. The resulting progenyfrom this cross are then crossed again to the recurrent parent and theprocess is repeated until a plant is obtained wherein essentially all ofthe desired morphological and physiological characteristics of therecurrent parent are recovered in the converted plant in addition to thetransferred gene(s) and associated trait(s) from the nonrecurrentparent.

Many gene traits have been identified that are not regularly selected inthe development of a new line but that can be improved by backcrossingtechniques. Gene traits may or may not be transgenic. Examples of thesetraits include, but are not limited to, male sterility, modified fattyacid metabolism, modified carbohydrate metabolism, herbicide resistance,pest or disease resistance (e.g., resistance to bacterial, fungal, orviral disease), insect resistance, enhanced nutritional quality,increased sweetness, increased flavor, improved ripening control,improved salt tolerance, industrial usage, yield stability, and yieldenhancement. These genes are generally inherited through the nucleus.

Tissue Culture.

Further reproduction of lettuce plants variety can occur by tissueculture and regeneration. Tissue culture of various tissues of lettuceand regeneration of plants therefrom is well known and widely published.For example, reference may be had to Teng, et al., HortScience, 27:9,1030-1032 (1992); Teng, et al., HortScience, 28:6, 669-1671 (1993);Zhang, et al., Journal of Genetics and Breeding, 46:3, 287-290 (1992);Webb, et al., Plant Cell Tissue and Organ Culture, 38:1, 77-79 (1994);Curtis, et al., Journal of Experimental Botany, 45:279, 1441-1449(1994); Nagata, et al., Journal for the American Society forHorticultural Science, 125:6, 669-672 (2000); and Ibrahim, et al., PlantCell Tissue and Organ Culture, 28(2), 139-145 (1992). It is clear fromthe literature that the state of the art is such that these methods ofobtaining plants are routinely used and have a very high rate ofsuccess. Thus, another aspect of this invention is to provide cellswhich upon growth and differentiation produce lettuce plants havingdesired characteristics of lettuce cultivar Solan. Optionally, lettuceplants can be regenerated from the tissue culture of the inventioncomprising all or essentially all of the physiological and morphologicalcharacteristics of lettuce cultivar Solan.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, suckers, and the like. Meansfor preparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and5,977,445 describe certain techniques.

Additional Breeding Methods.

This invention is also directed to methods for producing a lettuce plantby crossing a first parent lettuce plant with a second parent lettuceplant wherein the first or second parent lettuce plant is a plant oflettuce cultivar Solan. Further, both first and second parent lettucecan come from lettuce cultivar Solan. Thus, any of the followingexemplary methods using lettuce cultivar Solan are part of thisinvention: selfing, backcrosses, hybrid production, crosses topopulations, double haploid production, and the like. All plantsproduced using lettuce cultivar Solan as at least one parent are withinthe scope of this invention, including those developed from lettuceplants derived from lettuce cultivar Solan. Advantageously, lettucecultivar Solan can be used in crosses with other, different, lettuceplants to produce the first generation (F1) lettuce hybrid seeds andplants with desirable characteristics. The lettuce plants of theinvention can also be used for transformation where exogenous transgenesare introduced and expressed by the plants of the invention. Geneticvariants created either through traditional breeding methods or throughtransformation of the cultivars of the invention by any of a number ofprotocols known to those of skill in the art are intended to be withinthe scope of this invention.

The following describes exemplary breeding methods that may be used withlettuce cultivar Solan in the development of further lettuce plants. Onesuch embodiment is a method for developing lettuce cultivar Solanprogeny lettuce plants in a lettuce plant breeding program comprising:obtaining a plant, or a part thereof, of lettuce cultivar Solan,utilizing said plant or plant part as a source of breeding material, andselecting a lettuce cultivar Solan progeny plant with molecular markersin common with lettuce cultivar Solan and/or with some, all oressentially all of the morphological and/or physiologicalcharacteristics of lettuce cultivar Solan (see, e.g., Tables 1 to 15).In representative embodiments, the progeny plant has at least 1, 2, 3,4, 5, 6, 7, 8, 9, 10 or more of the morphological and physiologicalcharacteristics of lettuce cultivar Solan (e.g., as described in Tables1 to 15, such as blonde leaf color, high resistances to Downy Mildew(Bremia lactucae 16-35:EU and 5-9US), TBSV, Nasonovia ribisnigri biotypeNr: 0) and intermediate resistance to LMV strain Ls-1, or even all ofthe morphological and physiological characteristics of lettuce cultivarSolan so that said progeny lettuce plant is not significantly differentfor said traits than lettuce cultivar Solan, as determined at the 5%significance level when grown in the same environmental conditions;optionally, with the presence of one or more desired additional traits(e.g., male sterility, disease resistance, pest or insect resistance,herbicide resistance, and the like). Breeding steps that may be used inthe breeding program include pedigree breeding, backcrossing, mutationbreeding and/or recurrent selection. In conjunction with these steps,techniques such as RFLP-enhanced selection, genetic marker enhancedselection (for example, SSR markers) and/or and the making of doublehaploids may be utilized.

Another representative method involves producing a population of lettucecultivar Solan progeny plants, comprising crossing lettuce cultivarSolan with another lettuce plant, thereby producing a population oflettuce plants that, on average, derives at least 6.25%, 12.5%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%,97%, 98% or 99% of its alleles (i.e., TAC) from lettuce cultivar Solan,e.g., at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the geneticcomplement of lettuce cultivar Solan. One embodiment of this inventionis the lettuce plant produced by this method and that has obtained atleast 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles from lettucecultivar Solan, and optionally is the result of a breeding processcomprising one or two breeding crosses and one or more of selfing,sibbing, backcrossing and/or double haploid techniques in anycombination and any order. In embodiments, the breeding process does notinclude a breeding cross, and comprises selfing, sibbing, backcrossingand or double haploid technology. A plant of this population may beselected and repeatedly selfed or sibbed with a lettuce plant resultingfrom these successive filial generations. Another approach is to makedouble haploid plants to achieve homozygosity.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr and Walt, Principles of CultivarDevelopment, pp. 261-286 (1987). In embodiments, the inventionencompasses Solan progeny plants having a combination of at least 2, 3,4, 5, 6, 7, 8, 9, 10 or more of the characteristics as described hereinfor lettuce cultivar Solan, so that said progeny lettuce plant is notsignificantly different for said traits than lettuce cultivar Solan, asdetermined at the 5% significance level when grown in the sameenvironmental conditions. Using techniques described herein and thoseknown in the art, molecular markers may be used to identify said progenyplant as progeny of lettuce cultivar Solan. Mean trait values may beused to determine whether trait differences are significant, andoptionally the traits are measured on plants grown under the sameenvironmental conditions.

Progeny of lettuce cultivar Solan may also be characterized throughtheir filial relationship with lettuce cultivar Solan, as for example,being within a certain number of breeding crosses of lettuce cultivarSolan. A breeding cross is a cross made to introduce new genetics intothe progeny, and is distinguished from a cross, such as a self or a sibcross or a backcross to Solan as a recurrent parent, made to selectamong existing genetic alleles. The lower the number of breeding crossesin the pedigree, the closer the relationship between lettuce cultivarSolan and its progeny. For example, progeny produced by the methodsdescribed herein may be within 1, 2, 3, 4, 5 or more breeding crosses oflettuce cultivar Solan.

In representative embodiments, a lettuce plant derived from lettucecultivar Solan comprises cells comprising at least one set ofchromosomes derived from lettuce cultivar Solan. In embodiments, thelettuce plant or population of lettuce plants derived from lettucecultivar Solan comprises, on average, at least 6.25%, 12.5%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% of its alleles (i.e., TAC) from lettuce cultivar Solan, e.g.,at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the geneticcomplement of lettuce cultivar Solan, and optionally is the result of abreeding process comprising one or two breeding crosses and one or moreof selfing, sibbing, backcrossing and/or double haploid techniques inany combination and any order. In embodiments, the breeding process doesnot include a breeding cross, and comprises selfing, sibbing,backcrossing and or double haploid technology. In embodiments, thelettuce plant derived from lettuce cultivar Solan is one, two, three,four, five or more breeding crosses removed from lettuce cultivar Solan.

In representative embodiments, a plant derived from lettuce cultivarSolan is a double haploid plant, a hybrid plant or an inbred plant.

In embodiments, a hybrid or derived plant from lettuce cultivar Solancomprises a desired added trait. In representative embodiments, alettuce plant derived from lettuce cultivar Solan comprises all of themorphological and physiological characteristics of lettuce cultivarSolan (e.g., as described in Tables 1 to 15, for example, blonde leafcolor, high resistances to Downy Mildew (Bremia lactucae 16-35: EU and5-9US), TBSV and/or Nasonovia ribisnigri biotype Nr: 0) and/orintermediate resistance to LMV strain Ls-1. In embodiments, the lettuceplant derived from lettuce cultivar Solan comprises essentially all ofthe morphological and physiological characteristics of lettuce cultivarSolan (e.g., as described in Tables 1 to 15) in any combination, forexample, blonde leaf color, high resistance to Downy Mildew (Bremialactucae 16-35: EU and 5-9US), TBSV and/or Nasonovia ribisnigri biotypeNr: 0) and/or intermediate resistance to LMV strain Ls-1, with theaddition of a desired added trait.

Those skilled in the art will appreciate that any of the traitsdescribed above with respect to plant transformation methods can beintroduced into a plant of the invention (e.g., lettuce cultivar Solanand hybrid lettuce plants and other lettuce plants derived therefrom)using breeding techniques.

Genetic Analysis of Lettuce Cultivar Solan.

The invention further provides a method of determining a geneticcharacteristic of lettuce cultivar Solan or a progeny thereof, e.g., amethod of determining a genotype of lettuce cultivar Solan or a progenythereof. In embodiments, the method comprises detecting in the genome ofa Solan plant, or a progeny plant thereof, at least a first polymorphism(e.g., using nucleic acid amplification, nucleic acid sequencing and/orone or more molecular markers). To illustrate, in embodiments, themethod comprises obtaining a sample of nucleic acids from the plant anddetecting at least a first polymorphism in the nucleic acid sample.Optionally, the method may comprise detecting a plurality ofpolymorphisms (e.g., two or more, three or more, four or more, five ormore, six or more, eight or more or ten or more polymorphisms, etc.) inthe genome of the plant. In representative embodiments, the methodfurther comprises storing the results of the step of detecting thepolymorphism(s) on a computer readable medium. The invention furtherprovides a computer readable medium produced by such a method.

Deposit Information

Applicants have made a deposit of at least 2500 seeds of lettucecultivar Solan with the American Type Culture Collection (ATCC), 10801University Boulevard, Manassas, Va., 20110-2209 U.S.A. under ATCCDeposit No ______ on ______. This deposit of lettuce variety Solan willbe maintained in the ATCC depository, which is a public depository, fora period of 30 years, or 5 years after the most recent request, or forthe effective life of the patent, whichever is longer, and will bereplaced if any of the deposited seed becomes nonviable during thatperiod. Additionally, Applicants have satisfied all the requirements of37 C.F.R. §§ 1.801-1.809, including providing an indication of theviability of the samples. Access to this deposit will be made availableduring the pendency of this application to the Commissioner uponrequest. Upon the issuance of a patent on the variety, the variety willbe irrevocably and without restriction released to the public byproviding access to the deposit of at least 2500 seeds of the varietywith the ATCC. Applicants impose no restrictions on the availability ofthe deposited material from the ATCC; however, Applicants have noauthority to waive any restrictions imposed by law on the transfer ofbiological material or its transportation in commerce. Applicants do notwaive any infringement of its rights granted under this patent or underthe Plant Variety Protection Act (7 USC § 2321 et seq.).

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding.However, it will be apparent that certain changes and modifications suchas single gene modifications and mutations, somaclonal variants, variantindividuals selected from large populations of the plants of the instantinbred and the like may be practiced within the scope of the invention.

1. A seed of lettuce cultivar Solan, a representative sample of seedhaving been deposited under ATCC Accession No. PTA-126532.
 2. A plant oflettuce cultivar Solan, a representative sample of seed having beendeposited under ATCC Accession No. PTA-126532.
 3. A lettuce plant, or apart thereof, having all of the physiological and morphologicalcharacteristics of the lettuce plant of claim
 2. 4. A progeny lettuceplant of the plant of claim 2 that is within one breeding cross oflettuce cultivar Solan and comprises at least 50% of the alleles oflettuce cultivar Solan, wherein the progeny lettuce plant comprises ablonde leaf color and resistances to Bremia lactucae 16-35:EU and 5-9US,Lettuce Mosaic Virus strain Ls-1, and Nasonovia ribisnigri biotype Nr:0.
 5. A seed that produces the plant of claim
 4. 6. A plant part of thelettuce plant of claim
 2. 7. The plant part of claim 6, wherein theplant part is a leaf, pollen, an ovule, an anther, a root, or a cell. 8.A tissue culture of regenerable cells of the plant of claim
 2. 9. Alettuce plant regenerated from the tissue culture of claim 8 or a selfedprogeny thereof, wherein said lettuce plant comprises all of thephysiological and morphological characteristics of lettuce cultivarSolan.
 10. A processed product from the plant of claim 2, wherein theprocessed product comprises cut, sliced, ground, pureed, dried, canned,jarred, washed, packaged, frozen and/or heated leaves.
 11. A method ofproducing lettuce seed, the method comprising crossing the plant ofclaim 2 with itself or a second lettuce plant and harvesting theresulting seed.
 12. An F1 lettuce seed produced by the method of claim11.
 13. An F1 lettuce plant, or a meristem, leaf, pollen, cotyledon,hypocotyl, root, root tip, anther, flower, flower bud, pistil, ovule,shoot, stem, stalk, petiole, pith, capsule, scion, rootstock, or fruitthereof, produced by growing the F1 seed of claim
 12. 14. A doubledhaploid plant produced from the F1 lettuce plant of claim
 13. 15. Amethod for producing a seed of a lettuce plant derived from the plant ofclaim 2, the method comprising: (a) crossing a plant of lettuce cultivarSolan with a second lettuce plant; (b) allowing seed to form; (c)growing a plant from the seed of step (b) to produce a plant derivedfrom lettuce cultivar Solan; (d) selfing the plant of step (c) orcrossing it to a second lettuce plant to form additional lettuce seedderived from lettuce cultivar Solan; and (e) optionally repeating steps(c) and (d) one or more times to generate further derived lettuce seedfrom lettuce cultivar Solan, wherein in step (c) a plant is grown fromthe additional lettuce seed of step (d) in place of growing a plant fromthe seed of step (b).
 16. A seed produced by the method of claim 15,wherein the seed comprises at least 50% of the alleles of lettucecultivar Solan and is within one breeding cross of lettuce cultivarSolan, and wherein the seed produces a lettuce plant that comprises ablonde leaf color and resistances to Bremia lactucae 16-35: EU and5-9US, Lettuce Mosaic Virus strain Ls-1, and Nasonovia ribisnigribiotype Nr:
 0. 17. A plant, or part thereof, produced by growing theseed of claim
 16. 18. A method of vegetatively propagating the plant ofclaim 2, the method comprising: (a) collecting tissue capable of beingpropagated from a plant of lettuce cultivar Solan; (b) cultivating thetissue to obtain proliferated shoots; (c) rooting the proliferatedshoots to obtain rooted plantlets; and (d) optionally, growing plantsfrom the rooted plantlets.
 19. Lettuce plantlet or plant obtained by themethod of claim 18, wherein the lettuce plantlet or plant comprises allof the physiological and morphological characteristics of lettucecultivar Solan.
 20. A method of introducing a desired added trait intolettuce cultivar Solan, the method comprising: (a) crossing the plant ofclaim 2 with a lettuce plant that comprises a desired added trait toproduce F1 progeny; (b) selecting an F1 progeny that comprises thedesired added trait; (c) crossing the selected F1 progeny with lettucecultivar Solan to produce backcross progeny; (d) selecting a backcrossprogeny comprising the desired added trait; and (e) optionally repeatingsteps (c) and (d) one or more times to produce a further backcrossprogeny plant derived from lettuce cultivar Solan comprising a desiredadded trait and otherwise all of the physiological and morphologicalcharacteristics of lettuce cultivar Solan, wherein in step (c) theselected backcross progeny produced in step (d) is used in place of theselected F1 progeny of step (b).
 21. The method of claim 20, wherein thedesired added trait is male sterility, pest resistance, insectresistance, disease resistance, herbicide resistance, or any combinationthereof.
 22. A lettuce plant produced by the method of claim 20 or aselfed progeny thereof, wherein the lettuce plant is the backcrossprogeny of step (e) or a selfed progeny thereof and has the desiredadded trait and otherwise all of the physiological and morphologicalcharacteristics of lettuce cultivar Solan.
 23. (canceled)
 24. Seed thatproduces the plant of claim
 22. 25. A method of producing a plant oflettuce cultivar Solan comprising a desired added trait, the methodcomprising introducing a transgene conferring the desired trait into theplant of claim
 2. 26. A lettuce plant produced by the method of claim 25or a selfed progeny thereof, wherein the lettuce plant is a transformedplant of lettuce cultivar Solan and has the desired added trait.
 27. Aseed of the plant of claim 26, wherein the seed produces a transformedplant of lettuce cultivar Solan that comprises the transgene and has thedesired added trait.
 28. A method of determining a genotype of lettucecultivar Solan, the method comprising: (a) obtaining a sample of nucleicacids from the plant of claim 2; and (b) detecting a polymorphism in thenucleic acid sample.
 29. A method of producing a lettuce leaf, themethod comprising: (a) growing the lettuce plant according to claim 2 toproduce a lettuce leaf; and (b) harvesting the lettuce leaf.
 30. Amethod of producing a lettuce leaf, the method comprising: (a) growingthe lettuce plant according to claim 22 to produce a lettuce leaf; and(b) harvesting the lettuce leaf.